Constellation, A MorphoSys Company, Cambridge, MA, USA.
Methods Mol Biol. 2022;2529:477-490. doi: 10.1007/978-1-0716-2481-4_20.
Potent and highly selective small-molecule inhibitors are needed to unravel the biological complexities of histone methyltransferases and to reveal their therapeutic potential. A prerequisite to developing these inhibitors is the identification of validated chemical matter for initiating a medicinal chemistry campaign. For the most part, finding these initial starting points occurs through screening of large, unbiased compound libraries. The size and nature of these libraries, coupled with the complexities of the bisubstrate utilizing histone methyltransferases, necessitates that the primary screen and subsequent hit triage be carefully considered.In this chapter, using EZH2 as a representative example, we describe a screening and hit triage campaign that identified validated chemical matter allowing initiation of medicinal chemistry studies. Moreover, we discuss a cell-based assay to support lead identification and optimization. The approach described here entailing a mixture of biochemical, biophysical and cell-based assays should be applicable to identifying validated starting points for other histone methyltransferases.
需要有效且高度选择性的小分子抑制剂来揭示组蛋白甲基转移酶的生物学复杂性，并揭示其治疗潜力。开发这些抑制剂的前提是确定经过验证的化学物质，以启动药物化学研究。在大多数情况下，通过筛选大型、无偏见的化合物库来找到这些初始起点。这些库的大小和性质，加上双底物利用组蛋白甲基转移酶的复杂性，要求仔细考虑初始筛选和后续命中分类。在本章中，我们以 EZH2 为例，描述了一项筛选和命中分类活动，该活动确定了有效的化学物质，从而可以启动药物化学研究。此外，我们还讨论了一种基于细胞的测定方法，以支持先导化合物的鉴定和优化。这里描述的方法涉及生化、生物物理和基于细胞的测定方法的混合物，应该适用于确定其他组蛋白甲基转移酶的有效起始点。
